Polyvinylalcohol three-dimensional matrices for improved long-term dynamic culture of hepatocytes

Rat hepatocytes were seeded on three-dimensional highly porous polyvinylalcohol (PVA) and aminoethyl-modified polyvinylalcohol (AE-PVA) matrices. Hepatocytes were cultured under static and dynamic conditions. The three-dimensional matrices offered an improved extracellular microenvironment for long-term (5 days) maintenance of hepatocytes, compared to reference monolayer cultures on collagen. Cellular adhesion exceeded 80% with a viability superior to 70%. The preservation of albumin secretion after 5 days of culture was two times higher for static cultures on three-dimensional matrices (18% on PVA, 13% on AE-PVA) and three times higher for dynamic three-dimensional cultures (25% PVA and AE-PVA), compared to the static two-dimensional culture on collagen film (8%). The biotransformation of ammonia into urea was also maintained throughout the culture period. The addition of the aminoethyl function demonstrated no toxicity for the hepatocyte cultures. This function could be suitable eventually to further improve the hepatocyte culture system by linking more specific adhesion molecules on the polymer surface. This study demonstrated the efficiency of polyvinylalcohol as a three-dimensional matrix coupled to a perfusion culture system, which improves extracellular conditions for hepatocyte survival and promotes preservation of long-term hepatospecific functions.